EP0482076A1 - Stable vaccine compositions containing interleukins. - Google Patents
Stable vaccine compositions containing interleukins.Info
- Publication number
- EP0482076A1 EP0482076A1 EP19900911344 EP90911344A EP0482076A1 EP 0482076 A1 EP0482076 A1 EP 0482076A1 EP 19900911344 EP19900911344 EP 19900911344 EP 90911344 A EP90911344 A EP 90911344A EP 0482076 A1 EP0482076 A1 EP 0482076A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- interleukin
- vaccine composition
- antigen
- bacterial
- protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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Classifications
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- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
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- A61P31/12—Antivirals
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- A—HUMAN NECESSITIES
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- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55522—Cytokines; Lymphokines; Interferons
- A61K2039/55527—Interleukins
Definitions
- a substance that enhances the immunogenicity of an antigen with which it is injected is called an adjuvant.
- an adjuvant One of the more effective adjuvants is Freund's adjuvant, a water-in-oil emulsion. Freund's adjuvant is most effective when live or killed mycobacteria are suspended in the emulsion (Freund's complete adjuvant) along with antigen.
- Freund's adjuvant is most effective when live or killed mycobacteria are suspended in the emulsion (Freund's complete adjuvant) along with antigen.
- the intense, chronic inflammation that results around deposits of the emulsion precludes the use of the adjuvant in man. Emulsions lacking mycobacteria (incomplete Freund's adjuvant) are less irritating and have been used in man.
- Another type of adjuvant is a suspension of minerals on which antigen is adsorbed.
- lymphokines have been shown to have adjuvant activity thereby enhancing the immune response to an antigen.
- rhIL-2 recombinant human IL-2
- This composition was prepared and used
- Nakamura et al. demonstrated that interferon-gamma induced a two- to five-fold enhancement of antibody formation to several antigens. Nakamura et al., Nature 307: 381-382 (1984). Interleukins have also been shown to enhance an immune response to other antigens.
- This invention pertains to stable interleukincontaining vaccine compositions comprising a mixture of antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in suspension.
- the mixture can comprise a preservative.
- Interleukins such as interleukin-l ⁇ , interleukin-1 ⁇ , interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6 and interleukin-7 can be used as adjuvants in combination with antigen (particularly glycoconjugates) adsorbed onto a mineral suspension such as alum (e.g., aluminum hydroxide or phosphate) to modulate the immune response to the antigen.
- the vaccine compositions can be stored. Detailed Description of the Invention
- the vaccine compositions of this invention comprise an adjuvant amount of an interleukin in combination with the antigen adsorbed onto a mineral in suspension and a preservative.
- the mineral is alum (e.g., aluminum hydroxide or aluminum phosphate) which is suspended in an aqueous medium .
- the interleukin functions to modulate the immune response to the antigen, while the alum stabilizes the biological activity of the interleukin. In the absence of alum, interleukins have short half lives.
- the vaccine compositions of the present invention can be stored for periods of time which would otherwise result in the destabilization of the interleukin. Stabilization will greatly extend the permissible time for the manufacture, shipment and storage of the vaccine formulations prior to administering of the vaccine
- interleukins can be used. These include interleukin-1 ⁇ , interleukin-1 ⁇ , interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6, interleukin-7, or mixtures of these. Portions of interleukins having immunomodulating activity can also be used.
- the preferred interleukin is interleukin-2.
- Interleukin can be obtained from any suitable source. They can be produced by recombinant DNA methodology. For example, the genes encoding several human interleukins have been cloned and expressed in a variety of host systems, permitting the production of large quantities of pure human interleukin. Further, certain T lymphocyte lines produce high levels of interleukin thus providing a source of the interleukin.
- the preservative can be any pharmaceutically acceptable preservative. These include thimerosal, phenol, m-cresol, benzyl alcohol, methyl or ethyl paraben, and 2-phenoxyethanol.
- Interleukin can be used as adjuvant for many different types of antigens.
- the antigens can be particulate antigens such as bacteria, viruses and macrocomponents of cells and soluble antigens such as proteins, peptides, glycoproteins and carbohydrates.
- Antigens of particular interest are viral or bacterial antigens, allergens, auto-immunity related antigens, tumor-associated antigens, oncogene products, parasite antigens, fungal antigens or fragments of these.
- the antigens can be obtained from natural sources or they can be produced by recombinant DNA technology or other artificial means.
- bacterial antigens of interest are those associated with the human bacterial pathogens including, for example, typable and nontypable Haemophilus influen- zae, Escherichia coli, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus pyogenes, Branhamella catarrhalis , Vibrio cholerae, Corynebacteria
- Clostridium tetani Clostridium tetani.
- Some specific bacterial antigens include bacterial surface and outer membrane proteins (e.g. from Haemophilus influenzae, Neisseria).
- bacterial surface proteins e.g. the M protein from Streptococcus pyogenes.
- Viral antigens from pathogenic viruses include but are not limited to, human immunodeficiency virus (types I and II), human T-cell leukemia virus (types I, II and III), respiratory syncytial virus, hepatitis A, hepatitis B, hepatitis C, non-A and non-B hepatitis virus, herpes simplex virus (types I and II), cytomegalovirus, influenza virus, parainfluenza virus, poliovirus, rotavirus, coronavirus, rubella virus, measles virus, varicella, Epstein Barr virus, adenovirus, papilloma virus and yellow fever virus.
- human immunodeficiency virus types I and II
- human T-cell leukemia virus types I, II and III
- respiratory syncytial virus hepatitis A, hepatitis B, hepatitis C, non-A and non-B hepatitis virus
- herpes simplex virus
- Several specific viral antigens of these pathogenic viruses include the F protein (especially antigens containing the F peptide 283-315 described in W089/02935 entitled "Respiratory Syncytial Virus: Vaccines and Diagnostic Assays" by Paradiso, P. et al.) and the N and G proteins of respiratory syncytial virus (RSV), VP4 (previously known as VP3), VP6 and VP7 polypeptides of rotavirus, envelope glycoprotelns of human immunodeficiency virus, the S and pre-S antigens of hepatitis B and herpes glycoproteins B and D.
- F protein especially antigens containing the F peptide 283-315 described in W089/02935 entitled "Respiratory Syncytial Virus: Vaccines and Diagnostic Assays" by Paradiso, P. et al.
- RSV respiratory syncytial virus
- VP4 previously known as VP3
- capsular polymers are sugar containing polymers, such as polymers of sugars, sugar acids, amino sugars, polyhydric alcohols and sugar phosphates.
- Several capsular polymers and oligomers are useful as vaccines.
- the capsular polymers (CP) can be derived from many different types of bacteria. These types include
- Haemophilus influenzae Streptococcus species including pneumoniae (particularly serotypes 1, 4, 5, 6A, 6B, 9V, 14, 18C, 19F, and 23F) pyogenes and agalactiae,
- Ne is seria meningitidos (such as serogroup a, b and c), Klebsiella pneumoniae, Pseudomonas aeruginosa and
- Non-bacterial polymers can be derived from yeast and fungi, for example, Cryptococcus neoformans, or carbohydrate containing units found uniquely on cancer cells or those found associated with allergens.
- the antigens of this invention can be used to elicit an immune response to an antigen in a vertebrate (such as a mammalian host).
- the method comprises administering to the animal, an immunologically effective dose of a vaccine composition comprising a mixture of an antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in susp ens ion and an added p re s e rvative .
- the vaccine compositions are useful for the prevention of microbial infections.
- the antigens may be administered in a pharmaceutically acceptable vehicle, such as physiological saline, or ethanol polyols (such as glycerol or propylene glycol).
- the vaccine composition may optionally comprise other adjuvants, such as vegetable oils or emulsions thereof, surface active substances, e.g., hexadecylamine, octadecyl amino acid esters, octadecylamine, lysolecithin, dimethyl-dioctadecylammonium bromide, N,N-dicoctadecyl-N'-N'bis (2-hydroxyethyl-propane diamine), methoxyhexadecylglycerol, and pluronic polyols; polyamines, e.g., pyran, dextransulfate, poly IC, carbopol; peptides, e.g., muramyl dipeptide, dimethylgly
- tuftsin immune stimulating complexes (ISCOMS); oil emulsions; and mineral gels.
- ISCOMS immune stimulating complexes
- the antigens of this invention may also be incorporated into liposomes or ISCOMS. Supplementary active ingredients may also be employed.
- the vaccines can be administered to a human or animal in a variety of ways. These include intradermal, transdermal (such as by the use of slow release polymers), intramuscular, intraperitoneal, intravenous, subcutaneous, oral and intranasal routes of administration.
- the amount of antigen employed in such vaccine will vary depending upon the identity of the antigen employed. Adjustment and manipulation of established dosage ranges used with traditional carrier antigens for adaptation to the present vaccines is well within the ability of those skilled in the art.
- the vaccines of the present invention are intended for use in the treatment of both immature and adult warm-blooded animals, and in particular humans. Also, the use of the present composition is not limited to prophylactic application; therapeutic application are also contemplated (e.g., AIDS prophylaxis and therapy).
- the adjuvant action of interleukin has a number of important implications: the adjuvant action of the interleukin can increase the concentration of protective antibodies produced against the antigen in the vaccinated organism. As a result, effective (i.e., protective) vaccination can be achieved with a smaller quantity of antigen than would be normally required. This reduction in the required amount of antigen may lead to more widespread use of vaccines which are difficult or costly to prepare. This is especially true in the developing nations which have very limited health care budgets and which face epidemics of respiratory diseases, diarrheal diseases and malaria. It may also provide for safer vaccination when the antigen is toxic at the concentration normally required for effective immunization. By reducing the amount of antigen, the risk of toxic reaction is reduced.
- Interleukins by means of their immunomodulating activity, can help evoke a protective immune response against marginally or non-immunogenic antigens.
- vaccine compositions containing fragments of larger proteins, synthetic antigens or products of recombinant DNA technology may be made more potent by mixing them with interleukins.
- vaccination regimens call for the administration of antigen over a period of weeks or months in order to stimulate a "protective" immune response.
- a protective immune response is an immune response sufficient to protect the immunized host or subject from productive infection by a particular pathogen or pathogens to which the vaccine is directed.
- Interleukin when coadministered with antigen and adsorbed onto a mineral in suspension can accelerate the generation of a protective immune response. This may reduce the time course of effective vaccination
- vaccine formulations of this invention are sufficiently stable at 4oC to allow the manufacture, shipment and storage of the vaccine formulations.
- Example I Adjuvant Effect of rhIL-1 or rhIL-2 in
- a low dose (0.1 ⁇ g/mouse) dose of Haemophilus type b CRM conjugate (HbOC) was formulated with or without alum (100 ⁇ g/mouse) and with various concentrations of rhIL-2, rhIL-1 ⁇ or rhIL-1 ⁇ ranging from 1 ⁇ 10 2 - 5 ⁇
- Interleukins used in the preparation of the vaccines were either reconstituted on the day of formulation or were recovered from diluted stock (25 ⁇ g/ml) maintained at -70°C.
- Groups of Swiss-Webs ter mice (Taconic Farms,
- Germantown, NY were injected intramuscularly (I.M.) at weeks 0 and 2 with 0.1 ml of the vaccine formulations.
- Table 1 shows the results observed when rhIL-2 was administered as adjuvant in a mouse vaccination, both in the absence of alum [columns denoted (-)], and in combination with alum [columns denoted (+)].
- Antibody concentrations are expressed as ⁇ g/ml and were determined at weeks 2, 4, and 6, following the administration of 0.1 ⁇ g of the antigen.
- the antigen used in these studies was HbOC.
- Doses of rhIL-2 administered ranged from 1 ⁇ 10 3 - 1 ⁇ 10 5 units/mouse.
- rhIL-2 without alum does have a stimulatory effect on antibody production
- the antibody concentration present in the control samples was 0.42 ⁇ g/ml and 2.62 ⁇ g/ml.
- the correlation observed was that antibody concentration increased, as the rhIL-2 concentration decreased below 1 ⁇ 10 5 .
- the dose which appeared to stimulate the highest antibody production was approximately 1 ⁇ 10 4 units per mouse.
- Tables 2 and 3 show results obtained from immunizations in which rhIL-1 ⁇ and rhIL-1/3 were used as adjuvant, respectively.
- Table 2 presents data obtained from anti-PRP antibody determinations (expressed in ⁇ g/ml) at biweekly intervals post-immunization. Again, the experiments were conducted either with, (+), or without, (-), alum.
- rhIL-1 ⁇ In the presence of alum, rhIL-1 ⁇ also had a stimulating effect. Surprisingly, rhIL-1 ⁇ demonstrated increasing ability to stimulate the immune response as its concentration was decreased. The optimal adjuvant amount of rhIL-1 ⁇ , in the presence of alum, was approximately 1 x 10 3 . At such concentrations the antibody concentration was found to be 2-3 fold greater than the otherwise identical samples without (-) alum.
- mice 100 ⁇ g/mouse and with or without rhIL-2 or rhIL-1 ⁇ at 1 ⁇ 10 4 or 1 ⁇ 10 5 units/mouse.
- Groups of Swiss-Webs ter mice (5 animals per group) were immunized intramuscularly at weeks 0 and 2. Animals were bled as indicated in Table 4. Three doses of RSV F protein (1, 0.1 and 0.01 ⁇ g/mouse) were administered. Of these only 0.01 ⁇ g was suboptimal under the conditions employed.
- Table 1 shows the results of an experiment demonstrating the immune response to HbOC with rhIL-2 with and without alum.
- an HbOC based vaccine stimulated an antibody response of 4.7 ⁇ g/ml, after a single
- Such an antibody concentration is above the threshold level generally accepted as necessary for protection.
- HbOC antigen 2.5 ⁇ g/mouse
- 10 units of rhIL-2 were mixed with 10 units of rhIL-2 and adsorbed onto aluminum phosphate and stored at 4°C.
- Table 5 presents results observed in an in vitro stability assay.
- 5 ⁇ 10 3 CTLL-2 cells were cultured with various concentrations of rhIL-2 standard and HbOC vaccines. Cells were incubated in RPMI media containing 10% FBS at 37° for 24 hours and pulsed with 1.0 ⁇ Ci/well [ 3 H]-thymidine for 16 hours.
- the preparation was centrifuged to pellet the alum and the supernatent was assayed for rhIL-2 activity. It was determined that approximately 2/3 of the rhIL-2 is adsorbed onto alum. As shown in Table 5, when tested for rhIL-2 activity at time points following aluminum phosphate absorption, it was determined that the lymphokine maintained its activity for up to two weeks. TABLE 5
- rhIL-2 supt 4,460 ⁇ 205 5,716 ⁇ 211 13,460 ⁇ 1231 rhIL-2+50 ⁇ g
- mice were immunized with an HbOC/Alum/IL-2 vaccine which had been stored as described above.
- Four groups of two DBA/2 mice were immunized with 10 ⁇ g (protein) HbOC in CFA, Alum, rhIL-2/Alum, or rhIL-2 on 3 consecutive weeks. Lymph nodes were removed one week after injection and single cell suspension was obtained.
- 3 x 10 LNC were cultured with m i to gens and various concentrations of DT, CRM, and TT.
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Abstract
This invention pertains to interleukins-containing vaccine compositions, comprising a mixture of antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in suspension and a preservative. Preferably, the mineral is alum. The interleukin can modulate the protective immune response to an antigen, while the mineral stabilizes the biological activity of the interleukin.
Description
S TABLE VACC INE COMPOS I TIONS CONTAI N ING Background
It is often desirable to enhance the immunogenic potency of an antigen in order to obtain a stronger immune response in the organism being immunized and to strengthen host resistance to the antigen-bearing agent. A substance that enhances the immunogenicity of an antigen with which it is injected is called an adjuvant. One of the more effective adjuvants is Freund's adjuvant, a water-in-oil emulsion. Freund's adjuvant is most effective when live or killed mycobacteria are suspended in the emulsion (Freund's complete adjuvant) along with antigen. However, the intense, chronic inflammation that results around deposits of the emulsion precludes the use of the adjuvant in man. Emulsions lacking mycobacteria (incomplete Freund's adjuvant) are less irritating and have been used in man. Another type of adjuvant is a suspension of minerals on which antigen is adsorbed.
Certain lymphokines have been shown to have adjuvant activity thereby enhancing the immune response to an antigen. For example, Good et al. demonstrate the use of recombinant human IL-2 (rhIL-2) adsorbed on alum to enhance the immune response to a malaria related
antigen. This composition was prepared and used
immediately and stability over time was not ascertained. Good, M.F. et al., J. Immunol. 141:972-977 (1988).
Nakamura et al. demonstrated that interferon-gamma induced a two- to five-fold enhancement of antibody formation to several antigens. Nakamura et al., Nature
307: 381-382 (1984). Interleukins have also been shown to enhance an immune response to other antigens.
Nencioni et al., J. Immunol. 139: 800 - 804 (1987); Howard et al., EP285441. Summary of the Invention
This invention pertains to stable interleukincontaining vaccine compositions comprising a mixture of antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in suspension. The mixture can comprise a preservative. Interleukins, such as interleukin-lα, interleukin-1β, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6 and interleukin-7 can be used as adjuvants in combination with antigen (particularly glycoconjugates) adsorbed onto a mineral suspension such as alum (e.g., aluminum hydroxide or phosphate) to modulate the immune response to the antigen. The vaccine compositions can be stored. Detailed Description of the Invention
The vaccine compositions of this invention comprise an adjuvant amount of an interleukin in combination with the antigen adsorbed onto a mineral in suspension and a preservative. Preferably, the mineral is alum (e.g., aluminum hydroxide or aluminum phosphate) which is suspended in an aqueous medium .
The interleukin functions to modulate the immune response to the antigen, while the alum stabilizes the biological activity of the interleukin. In the absence of alum, interleukins have short half lives. Thus, the vaccine compositions of the present invention can be
stored for periods of time which would otherwise result in the destabilization of the interleukin. Stabilization will greatly extend the permissible time for the manufacture, shipment and storage of the vaccine formulations prior to administering of the vaccine
formulation.
Several different interleukins can be used. These include interleukin-1α, interleukin-1β, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6, interleukin-7, or mixtures of these. Portions of interleukins having immunomodulating activity can also be used. The preferred interleukin is interleukin-2.
Interleukin can be obtained from any suitable source. They can be produced by recombinant DNA methodology. For example, the genes encoding several human interleukins have been cloned and expressed in a variety of host systems, permitting the production of large quantities of pure human interleukin. Further, certain T lymphocyte lines produce high levels of interleukin thus providing a source of the interleukin.
The preservative can be any pharmaceutically acceptable preservative. These include thimerosal, phenol, m-cresol, benzyl alcohol, methyl or ethyl paraben, and 2-phenoxyethanol.
Interleukin can be used as adjuvant for many different types of antigens. In general, the antigens can be particulate antigens such as bacteria, viruses and macrocomponents of cells and soluble antigens such as proteins, peptides, glycoproteins and carbohydrates. Antigens of particular interest are viral or bacterial antigens, allergens, auto-immunity related antigens,
tumor-associated antigens, oncogene products, parasite antigens, fungal antigens or fragments of these. The antigens can be obtained from natural sources or they can be produced by recombinant DNA technology or other artificial means.
Among the bacterial antigens of interest are those associated with the human bacterial pathogens including, for example, typable and nontypable Haemophilus influen- zae, Escherichia coli, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus pyogenes, Branhamella catarrhalis , Vibrio cholerae, Corynebacteria
diphtheriae, Neisseria gonorrhoeae, Bordetella pertussis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes, Klebsiella pneumoniae, and
Clostridium tetani. Some specific bacterial antigens include bacterial surface and outer membrane proteins (e.g. from Haemophilus influenzae, Neisseria
meningitidis, Neisseria gonorrhoeae or Branhamella catarrhalis) and bacterial surface proteins (e.g. the M protein from Streptococcus pyogenes).
Viral antigens from pathogenic viruses include but are not limited to, human immunodeficiency virus (types I and II), human T-cell leukemia virus (types I, II and III), respiratory syncytial virus, hepatitis A, hepatitis B, hepatitis C, non-A and non-B hepatitis virus, herpes simplex virus (types I and II), cytomegalovirus, influenza virus, parainfluenza virus, poliovirus, rotavirus, coronavirus, rubella virus, measles virus, varicella, Epstein Barr virus, adenovirus, papilloma virus and yellow fever virus.
Several specific viral antigens of these pathogenic viruses include the F protein (especially antigens containing the F peptide 283-315 described in W089/02935 entitled "Respiratory Syncytial Virus: Vaccines and Diagnostic Assays" by Paradiso, P. et al.) and the N and G proteins of respiratory syncytial virus (RSV), VP4 (previously known as VP3), VP6 and VP7 polypeptides of rotavirus, envelope glycoprotelns of human immunodeficiency virus, the S and pre-S antigens of hepatitis B and herpes glycoproteins B and D.
Also of interest are various antigens associated with auto-immune diseases, such as rheumatoid arthritis and lupus erythematosus.
Of particular interest for use in a vaccine are capsular polymers (CP) produced by bacterial pathogens. Capsular polymers are sugar containing polymers, such as polymers of sugars, sugar acids, amino sugars, polyhydric alcohols and sugar phosphates. Several capsular polymers and oligomers are useful as vaccines.
The capsular polymers (CP) can be derived from many different types of bacteria. These types include
Haemophilus influenzae, Streptococcus species including pneumoniae (particularly serotypes 1, 4, 5, 6A, 6B, 9V, 14, 18C, 19F, and 23F) pyogenes and agalactiae,
Ne is seria meningitidos (such as serogroup a, b and c), Klebsiella pneumoniae, Pseudomonas aeruginosa and
Staphylococcus aureus.
Non-bacterial polymers can be derived from yeast and fungi, for example, Cryptococcus neoformans, or carbohydrate containing units found uniquely on cancer cells or those found associated with allergens.
The antigens of this invention can be used to elicit an immune response to an antigen in a vertebrate (such as a mammalian host). The method comprises administering to the animal, an immunologically effective dose of a vaccine composition comprising a mixture of an antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in susp ens ion and an added p re s e rvative . The vaccine compositions are useful for the prevention of microbial infections. The antigens may be administered in a pharmaceutically acceptable vehicle, such as physiological saline, or ethanol polyols (such as glycerol or propylene glycol). The vaccine composition may optionally comprise other adjuvants, such as vegetable oils or emulsions thereof, surface active substances, e.g., hexadecylamine, octadecyl amino acid esters, octadecylamine, lysolecithin, dimethyl-dioctadecylammonium bromide, N,N-dicoctadecyl-N'-N'bis (2-hydroxyethyl-propane diamine), methoxyhexadecylglycerol, and pluronic polyols; polyamines, e.g., pyran, dextransulfate, poly IC, carbopol; peptides, e.g., muramyl dipeptide, dimethylglycine,
tuftsin; immune stimulating complexes (ISCOMS); oil emulsions; and mineral gels. The antigens of this invention may also be incorporated into liposomes or ISCOMS. Supplementary active ingredients may also be employed.
The vaccines can be administered to a human or animal in a variety of ways. These include intradermal, transdermal (such as by the use of slow release polymers), intramuscular, intraperitoneal, intravenous,
subcutaneous, oral and intranasal routes of administration. The amount of antigen employed in such vaccine will vary depending upon the identity of the antigen employed. Adjustment and manipulation of established dosage ranges used with traditional carrier antigens for adaptation to the present vaccines is well within the ability of those skilled in the art. The vaccines of the present invention are intended for use in the treatment of both immature and adult warm-blooded animals, and in particular humans. Also, the use of the present composition is not limited to prophylactic application; therapeutic application are also contemplated (e.g., AIDS prophylaxis and therapy).
The adjuvant action of interleukin has a number of important implications: the adjuvant action of the interleukin can increase the concentration of protective antibodies produced against the antigen in the vaccinated organism. As a result, effective (i.e., protective) vaccination can be achieved with a smaller quantity of antigen than would be normally required. This reduction in the required amount of antigen may lead to more widespread use of vaccines which are difficult or costly to prepare. This is especially true in the developing nations which have very limited health care budgets and which face epidemics of respiratory diseases, diarrheal diseases and malaria. It may also provide for safer vaccination when the antigen is toxic at the concentration normally required for effective immunization. By reducing the amount of antigen, the risk of toxic reaction is reduced.
Interleukins, by means of their immunomodulating activity, can help evoke a protective immune response against marginally or non-immunogenic antigens. In this manner, vaccine compositions containing fragments of larger proteins, synthetic antigens or products of recombinant DNA technology may be made more potent by mixing them with interleukins.
Typically, vaccination regimens call for the administration of antigen over a period of weeks or months in order to stimulate a "protective" immune response. A protective immune response, is an immune response sufficient to protect the immunized host or subject from productive infection by a particular pathogen or pathogens to which the vaccine is directed. Interleukin, when coadministered with antigen and adsorbed onto a mineral in suspension can accelerate the generation of a protective immune response. This may reduce the time course of effective vaccination
regimens. In some instances, it may result in the generation of a protective immune response in a single dosage. Further, vaccine formulations of this invention are sufficiently stable at 4ºC to allow the manufacture, shipment and storage of the vaccine formulations.
This invention is further illustrated by the following examples.
EXAMPLES
Example I: Adjuvant Effect of rhIL-1 or rhIL-2 in
the Presence of Alum With an HbOC
Antigen Vaccine
A low dose (0.1 μg/mouse) dose of Haemophilus type b CRM conjugate (HbOC) was formulated with or without
alum (100 μg/mouse) and with various concentrations of rhIL-2, rhIL-1α or rhIL-1β ranging from 1 × 102 - 5 ×
105 units/mouse. All vaccines were prepared on the day of immunization and maintained at 4ºC until injected.
Interleukins used in the preparation of the vaccines were either reconstituted on the day of formulation or were recovered from diluted stock (25 μg/ml) maintained at -70°C. Groups of Swiss-Webs ter mice (Taconic Farms,
Germantown, NY) were injected intramuscularly (I.M.) at weeks 0 and 2 with 0.1 ml of the vaccine formulations.
Serum samples were collected as indicated in the various
Tables. A. rhIL-2 as Adjuvant
Table 1 shows the results observed when rhIL-2 was administered as adjuvant in a mouse vaccination, both in the absence of alum [columns denoted (-)], and in combination with alum [columns denoted (+)]. Antibody concentrations are expressed as μg/ml and were determined at weeks 2, 4, and 6, following the administration of 0.1 μg of the antigen. The antigen used in these studies was HbOC. Doses of rhIL-2 administered ranged from 1 × 103 - 1 × 105 units/mouse.
TABLE 1
HbOC Units Week 2 Week 4 Week 6 μE rhIL (+) (-) (+) (-) (+) (-)
Series 1 :
0.1 103 4.70 0.28 7.89 3.46 8.16 3.62 0.1 104 2.25 0.51 9.39 5.01 17.77 5.83
0.1 105 0.37 0.55 4.71 3.04 5.31 2.91
0.1 - - 0.46 <0.10 3.53 0.42 5.46 3.53
Series 2
0.1 102 1.02 0.40 6.32 4.21 5.90 3.32
0.1 103 0.54 0. .75 5.77 4.73 5.60 4.61
0.1 104 1.68 <0. .10 12.99 1.95 19.06 1.25
0.1 105 0.25 <0, .10 5.94 5.46 3.91 3.82
0.1 - - 0.53 0. .30 7.09 2.64 8.30 2.82 As can be seen in the columns denoted (-) above, rhIL-2 without alum does have a stimulatory effect on antibody production For example, in two separate experiments, the antibody concentration present in the control samples was 0.42 μg/ml and 2.62 μg/ml.
The results observed when alum was included with rhIL-2 in the adjuvant mixture are shown in the columns denoted (+) in Table 1. Again, a significant stimulatory effect is seen. However, when alum is present, the magnitude of the antibody response is significantly increased over the response observed with otherwise identical samples without alum (vaccinations).
Again considering the 4 week time point, it can be seen that the with (+) alum antibody concentrations are
up to more than 10-fold greater than the corresponding without (-) alum antibody concentrations. Furthermore, there is an apparent rhIL-2 concentration dependence which was absent in the samples without (-) alum.
Specifically, the correlation observed was that antibody concentration increased, as the rhIL-2 concentration decreased below 1 × 105. In the case of rhIL-2 with alum, the dose which appeared to stimulate the highest antibody production was approximately 1 × 104 units per mouse.
B. rhIL-1 as Adjuvant
Tables 2 and 3 show results obtained from immunizations in which rhIL-1 α and rhIL-1/3 were used as adjuvant, respectively. Table 2 presents data obtained from anti-PRP antibody determinations (expressed in μg/ml) at biweekly intervals post-immunization. Again, the experiments were conducted either with, (+), or without, (-), alum.
TABLE 2
HbOC rhlL-1α Week 2 Week 4 Week 6 μg (Units) (+) (-) (+) (-) (+) (-)
Series 1:
0.1 103 1.96 1.02 14.63 4.94 13.25 4.93
0.1 104 0.70 2.54 7.16 5.11 12.44 11.17
0.1 105 0.33 1.49 4.69 7.02 6.14 8.73
0.1 -IL 0.46 <0.10 3.53 0.42 5.46 3.53 Control Series 2
0.1 102 <0.10 1.02 3.68 4.36 4. .07 3. .00
0.1 103 1.06 1.17 12.31 5.54 8. .74 3. .08
0.1 104 0.47 1.47 4.73 8.97 5. .98 6. .55
0.1 105 0.34 1.00 8.55 16.74 7 .77 16, .29
0.1 -IL 0.53 0.30 7.09 2.64 8 .30 2 .82 Control
The results observed from rhIL-1α and rhIL-1β adjuvant mixtures were similar, overall, to those observed in the rhIL-2 series. Table 2, for example, shows that when rhIL-1α Is administered as adjuvant without alum there is a stimulatory effect when compared with an otherwise identical vaccine without the lymphokine. As shown in the columns denoted (-), a tendency toward decreasing antibody production was observed as the amount of rhIL-1α in the adjuvant mixture was decreased from 1 × 105 to 1 × 102 units per mouse.
In the presence of alum, rhIL-1α also had a stimulating effect. Surprisingly, rhIL-1α demonstrated
increasing ability to stimulate the immune response as its concentration was decreased. The optimal adjuvant amount of rhIL-1α, in the presence of alum, was approximately 1 x 103. At such concentrations the antibody concentration was found to be 2-3 fold greater than the otherwise identical samples without (-) alum.
Similar results were observed when rhIL-1β was used as adjuvant, as shown in Table 3. The data in Table 3 summarizes the results of antibody concentration determinations made at weeks 2, 4, and 6, for varying concentrations of rhIL-1β as adjuvant, either with or without alum. In all but 6 of 21 experimental groups, the vaccine with (+) alum resulted in a higher antibody concentration than the vaccine without (-) alum.
TABLE 3
HbOC rhlL- 1β Week 2 Week 4 Week 6 μg (+) (-) (+) (-) (+) (-)
Series 1:
0.1 103 0.41 0.96 5.27 4.09 8.32 5.22
0.1 104 0.90 0.62 4.13 3.39 8.15 6.39
0.1 105 1.46 0.46 13.35 1.77 8.60 1.06
0.1 -IL 0..46 <0.10 3.53 0.42 5.46 3.53
Control Series 2:
0.1 102 0. .85 0.16 8.76 0.04 6. ,66 1.91
0.1 103 0. .30 0.40 4.57 0.68 6. .87 10.33
0.1 104 1. .08 0.36 5.75 5.15 5. .17 4.47
0.1 105 0. .19 0.36 2.34 3.0 4. .98 3.92
0.1 -IL 0. .53 0.30 7.09 2.64 8. .30 2.82 Control
Example II : Adjuvant Effect of rhIL-1α, rhlL-1β,
rhIL -2, and Mixtures Thereof on an RSV F Protein Vaccine
To determine whether interleukins can be used to enhance the antibody response to a protein vaccine, various concentrations of F protein of respiratory syncytial virus (RSV) were formulated with alum (final
100 μg/mouse) and with or without rhIL-2 or rhIL-1α at 1 × 104 or 1 × 105 units/mouse. Groups of Swiss-Webs ter mice (5 animals per group) were immunized intramuscularly at weeks 0 and 2. Animals were bled as indicated in Table 4.
Three doses of RSV F protein (1, 0.1 and 0.01 μg/mouse) were administered. Of these only 0.01 μg was suboptimal under the conditions employed. Comparison of the response seen in those groups receiving vaccines containing various interleukins with the control group receiving 0.01 μg of protein in alum alone revealed no significant effect (4 fold difference in titer) of rhIL-2 or rhlL-1α on the antibody response to F protein.
However, the responses in the rhIL-2 treated groups were higher than controls at doses of 1 × 104 units/mouse which is similar to the results obtained in the HbOC studies. With IL-1α, both doses seem to show some improvement in antibody responses over controls.
Interestingly, mixtures of 1 × 104 units/mouse of rhIL-2 and rhIL-1α did not show any indication of synergy but rather showed a slight decrease in the response relative to controls suggesting a possible antagonistic interaction of the interleukins.
TABLE 4
Adjuvant Effect of rhIL-2 on RSV F protein vaccine
μgF protein rhIL Units Week 2 Week 4 Week 6
0. 1 -IL
Control - - 141,657 2,341,756 2,454,512
0. 1 IL-2 104 191,935 2,045,327 2,101,951
0. 1 IL-2 105 309,797 2,275,311 2,311,326
0. 01 -IL
Control - - 68,627 802,611 687,334
0. 01 IL-2 104 145,467 1,580,552 1,699,135
0. 01 IL-2 105 60,134 722,396 815,351
0.1 -IL
Control - - 141,657 2,341,756 2,454,512
0.1 IL-1α 104 123,446 1,861,917 1,771,952
0.1 IL-1α 105 79,386 1,185,475 1,214,008
0.01 -IL
Control - - 68,627 802,611 687,334
0 .01 IL-1α 104 54,081 1,003,110 1,094,459
0 .01 IL-1α 105 59,566 708,009 1,178,056
0 .1 -IL
Control - - 141,657 2,341,756 2,451,512
0 .1 Mix 104 207,410 2,593,957 2,353,912
0 .01 -IL
Control - - 68,627 802,611 687,334
0 .01 Mix 104 15,947 367,224 532,050
1 .0 - - 211,662 2,269,615 2,899,079
Example III. Single Dose Vaccination
Table 1 shows the results of an experiment demonstrating the immune response to HbOC with rhIL-2 with and without alum. At a concentration of 1 × 103 units rhIL-2, in the presence of alum, an HbOC based vaccine stimulated an antibody response of 4.7 μg/ml, after a single
administration. Such an antibody concentration is above the threshold level generally accepted as necessary for protection.
Example IV Adjuvant Stability
A . ln Vitro Assay of rhIL-2 Stability
To study the stability of rhIL-2 in an alum containing composition, HbOC antigen (2.5 μg/mouse) was mixed with 10 units of rhIL-2 and adsorbed onto aluminum phosphate and stored at 4°C. Table 5 presents results observed in an in vitro stability assay. In the assay, 5 × 103 CTLL-2 cells were cultured with various concentrations of rhIL-2 standard and HbOC vaccines. Cells were incubated in RPMI media containing 10% FBS at 37° for 24 hours and pulsed with 1.0 μCi/well [3H]-thymidine for 16 hours. To estimate the amount of rhIL-2 adsorbed to alum, the preparation was centrifuged to pellet the alum and the supernatent was assayed for rhIL-2 activity. It was determined that approximately 2/3 of the rhIL-2 is adsorbed onto alum. As shown in Table 5, when tested for rhIL-2 activity at time points following aluminum phosphate absorption, it was determined that the lymphokine maintained its activity for up to two weeks.
TABLE 5
In vitro assay of rhIL-2 stability (Incorporation of
[3H]-thymidine at 1.0 U/ml IL-2 A ±CPM ± SD).
Week 0 Week 1 Week 2
Baseline:
Media
(× cpm) 303 ± 41 1,540 ± 67 135 ± 49
Interleukin-2:
BM 16,179 ± 491 10,056 ± 689 13,323 ± 1428 product
Cetus 28,300 ± 2250 25,681 ± 135 25,992 ± 868 product
HbOC vaccines:
rhIL-2 24,188 ± 783 31,989 ± 2252 36,312 ± 3102 Alum/
rhIL-2 19,419 ± 1330 28,785 ± 1691 35,091 ± 690 Alum/
rhIL-2 supt 4,460 ± 205 5,716 ± 211 13,460 ± 1231 rhIL-2+50μg
Alum ND ND 35,627 ± 2503
B. In Vivo Assay of rhIL-2 Stability
An in vivo assay of rhIL-2 stability was designed. Mice were immunized with an HbOC/Alum/IL-2 vaccine which had been stored as described above. Four groups of two DBA/2 mice were immunized with 10μg (protein) HbOC in CFA, Alum, rhIL-2/Alum, or rhIL-2 on 3 consecutive weeks. Lymph nodes were removed one week after injection and single cell
suspension was obtained. 3 x 10 LNC were cultured with m i to gens and various concentrations of DT, CRM, and TT. Cells were incubated in RPMI media containing 1.0% NMS at 37°C for 3 days, pulsed with 1.0 μCi/well [3H]-thymidine for 16 hrs, and harvested for counting on LS counter. A significant T cell response was observed in the HbOC-CFA group. In weeks 2 and 3, however, rhIL-2 induced an augmented T cell response. Furthermore, even HbOC alone appears to protect rhIL-2 from degradation. Equivalents
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims:
TABLE 6
In Vivo Stability of IL-2 with HbOC Vaccine
Maximum Incorporated [3H]-thymidine as SI ± SD
Week 1 HbOC Priming In:
In Vitro
Challenge: CFA Alum* IL-2/Alum* IL-2*
Media (× cpm) 454 ± 22 190 ±19 386 ±35 125 ± 8
CA 1.0μg/ml 43.3± 3.9 89.7± 3.9 58.4± 5.1 136. 9± 2. 4
LPS 50μg/ml 129.4± 8.7 189.9± 7.7 69.0± 4.5 103. 9± 6. 6
DT 17.5± 1.2 4.4± 0.04 1.4± 0.3 1. 8± 0. 1
CRM 162.0± 14.9 9.8± 0.3 0.8± 0.1 0. 8± 0. 1
TT 0.8± 0.07 0.4± 0.03 0.1± 0.0 0. 6± 0. 05
Week 2
Media 1,185 ±175 590 ±42 552 ±48 187 ±11
CA 1.0μg/ml 40.6± 3.4 57.5± 1.8 53.1± 0.9 165.1+11.5
LPS 50μg/ml 71.5± 1.2 89.9± 3.7 142.1± 8.7 203.5±17.8
DT 16.5± 0.4 1.1± 0.0 1.3± 0.2 2.9± 0.0
CRM 50.6± 0.5 1.3± 0.0 10.5± 0.3 1.6± 0.0
TT 0.6± 0.1 0.1± 0.0 0.1± 0.0 0.4± 0.0
Week 3
Media 519 ± 59 416 ± 9 442 ±63 966 ±51
CA 1.0μg/ml 45.9± 0.3 41.0± 2.8 46.4± 0.9 27 .5± 2 .1
LPS 50μg/ml 129.2± 7.6 119.4± 5.3 126.5± 6.3 81 .9± 3 .1
DT 5.6± 0.8 NA 2.2± 0.2 NA
CRM 98.1± 8.1 17.9± 1.4 25.2± 1.6 31 .4± 2 .6
TT 0.4± 0.1 0.2± 0.0 0.51 0.0 0.1± 0 .0
*These groups received 2.0% NMS instead of 1.0%
Claims
1. A stable vaccine composition, comprising a mixture of an antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in suspension and a pharmaceutically acceptable preservative.
2. The vaccine composition of Claim 1, wherein the interleukin is selected from the group consisting of interleukin-1α, interleukin-1β, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6, interleukin-7, or mixtures thereof.
The vaccine composition of Claim 2, wherein the interleukin is human interleukin-2.
4. The vaccine composition of Claim 1, wherein the mineral suspension is an aqueous suspension of alum.
5. The vaccine composition of Claim 1, wherein the antigen is an antigen selected from the group consisting of bacteria, viruses, macro - components of cells, proteins, peptides, glycoproteins, carbohydrates, parasites, fungi, oncogene products and cancer cells.
6. A vaccine composition of Claim 5, wherein the
bacterial antigen is from a bacterial pathogen,
7. The vaccine composition of Claim 6, wherein the bacterial pathogen is selected from the group consisting of Haemophilus influenzae, Escherichia coli, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus pyogenes, Branhamella catarrhalis, Vibrio cholerae, Corynebacteria diphtheriae, Neisseria gonorrhoeae, Bordetella pertussis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes, Klebsiella
pneumoniae and Clostridium tetani.
8. The vaccine composition of Claim 1, wherein the antigen is a bacterial capsular polymer, oligmer, or fragment thereof.
9. The vaccine composition of Claim 8, wherein the bacterial capsular polymer, oligomer or fragment thereof is Haemophilus influenzae, Streptococcus pheumoniae, Neisseria meningitidis, Klebsiella pneumoniae, Pseudomonas aeruginosa or Staphylococcus aureus.
10. The vaccine composition of Claim 1, wherein the antigen is coupled to a glycoconjugate.
11. The vaccine composition of Claim 10, wherein the glycoconjugate comprises a bacterial toxin of diphtheria, tetanus, pertussis or CRM, or toxoid thereof.
12. The vaccine composition of Claim 10, wherein the glycoconjugate comprises polyribosylribitolphosphate and CRM197 of diphtheria toxin.
13. A vaccine composition of Claim 1, wherein the
bacterial antigen is a bacterial surface or outer membrane protein.
14. The vaccine composition of Claim 13, wherein the bacterial surface or outer membrane protein is of Haemopfilus influenzae, Neisseria meningitidis, Neisseria gonorrhoea or Branhamella catarrhalis.
15. A vaccine composition of Claim 13, wherein the
bacterial surface protein is the M protein from
Streptococcus pyogenes.
16. A vaccine composition of Claim 5, wherein the viral antigen is selected from the group consisting of F protein of respiratory syncytial virus, N protein of respiratory syncytial virus, G protein of respiratory syncytial virus, VP4 polypeptide of rotavirus, VP6 polypeptide of rotavirus, VP7 polypeptide of rotavirus, envelope glycoproteins of human immunodeficiency virus, herpes glycoproteins B and D and the S and pre-S antigens of hepatitis B.
17. A vaccine composition of Claim 1 wherein the
preservative is thimerosal, phenol, benzyl alcohol, methyl or ethyl paraben, 2-phenoxyethanol or m-cresol.
18. A stable vaccine composition comprising, a mixture of an antigen and an adjuvant amount of interleuken-1α, interleukin-1β, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6 or interleukin-7, adsorbed onto a mineral in suspension.
19. A vaccine composition of Claim 18, further comprising a preservative.
20. A vaccine composition of Claim 19, wherein the
preservative is thimerosal, phenol, m-cresol, benzyl alcohol, methyl or ethyl paraben, or 2-phenoxyethanol.
21. The vaccine composition of Claim 18, wherein the mineral suspension is an aqueous suspension of alum.
22. The vaccine composition of Claim 18, wherein the antigen is an antigen selected from the group consisting of bacteria, viruses, macro-components of cells, proteins, peptides, glycoproteins, carbohydrates, parasites, fungi, oncogene products and cancer cells.
23. A vaccine composition of Claim 22, wherein the
bacterial antigen is from a bacterial pathogen.
24. The vaccine composition of Claim 23, wherein the bacterial pathogen is selected from the group consisting of Haempphilus influenzae, Escherichia coli, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus pyogenes, Branhamella
catarrhalis, Vibrio cholerae, Corynebacteria diphtheriae, Neisseria gonorrhoeae, Bordetella pertussis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes, Klebsiella
pneumoniae and Clostridium tetani.
25. The vaccine composition of Claim 18, wherein the antigen is a bacterial capsular polymer, oligmer or fragment thereof.
26. The vaccine composition of Claim 25, wherein the bacterial capsular polymer, oligomer or fragment thereof is Haemophilus influenzae, Streptococcus pheumoniae, Neisseria meningitidis, Klebsiella pneumoniae, Pseudomonas aeruginosa or Staphylococcus aureus.
27. The vaccine composition of Claim 18, wherein the antigen is coupled to a glycoconjugate.
28. The vaccine composition of Claim 27, wherein the glycoconjugate comprises a bacterial toxin of diphtheria, tetanus or pertussis, or CRM or toxoid thereof.
29. The vaccine composition of Claim 28, wherein the glycoconjugate comprises polyribosylribitolphosphate and CRM197 of diphtheria toxin.
30. A vaccine composition of Claim 18, wherein the
bacterial antigen is a bacterial surface or outer membrane protein.
31. The vaccine compo s iti on o f Claim 30 , where in the bac terial surface or outer membrane protein is of Haemoplilus influenzae, Neisseria meningitidis,
Neisseria gonorrhoeae or Branhamella catarrhalis.
32. A vaccine composition of Claim 30, wherein the
bacterial surface protein is the M protein from
Streptococcus pyogenes.
33. A vaccine composition of Claim 22, wherein the
viral antigen is selected from the group consisting of F protein of respiratory syncytial virus, N protein of respiratory syncytial virus, G protein of respiratory syncytial virus, VP4 polypeptide of rotavirus, VP6 polypeptide of rotavirus, VP7 polypeptide of rotavirus, envelope glycoproteins of human immunodeficiency virus, herpes glycoprotein B and D and the S and pre-S antigens of hepatitis B.
34. A method of eliciting an immune response against an antigen, comprising administering to a vertebrate host an effective amount of a stable vaccine composition, comprising a mixture of an antigen and an adjuvant amount of an interleukin adsorbed onto a mineral in suspension and a pharmaceutically acceptable preservative, in a pharmaceutically acceptable vehicle and optional adjuvant.
35. The method of Claim 34, wherein the interleukin is selected from the group consisting of interleukin-1α, interleukin-1β, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-6, interleukin 7 or mixtures thereof.
36. The method of Claim 34, wherein the mineral suspension is an aqueous suspension of alum.
37. The method of Claim 34, wherein the antigen is an antigen selected from the group consisting of bacteria, viruses, macro-components of cells, proteins, peptides, glycoproteins, carbohydrates, parasites, fungi, oncogene products and cancer cells.
38. A method of eliciting an immune response against an antigen, comprising administering to a vertebrate host an effective amount of a stable vaccine composition comprising, a mixture of an antigen and an adjuvant amount of interleuken-1α, interleukin-1β, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin- 6 or interleukin-7 adsorbed onto a mineral in suspension, in a pharmaceutically acceptable vehicle and optional adjuvant.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
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| US37974289A | 1989-07-14 | 1989-07-14 | |
| US379742 | 1989-07-14 | ||
| PCT/US1990/003982 WO1991001143A1 (en) | 1989-07-14 | 1990-07-16 | Stable vaccine compositions containing interleukins |
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| Publication Number | Publication Date |
|---|---|
| EP0482076A1 true EP0482076A1 (en) | 1992-04-29 |
| EP0482076B1 EP0482076B1 (en) | 1995-04-26 |
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ID=23498494
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP90911344A Expired - Lifetime EP0482076B1 (en) | 1989-07-14 | 1990-07-16 | Stable vaccine compositions containing interleukins |
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| EP (1) | EP0482076B1 (en) |
| JP (2) | JP3485184B2 (en) |
| KR (1) | KR0177179B1 (en) |
| AT (1) | ATE121629T1 (en) |
| AU (1) | AU648509B2 (en) |
| CA (1) | CA2063587C (en) |
| DE (1) | DE69018990T2 (en) |
| DK (1) | DK0482076T3 (en) |
| ES (1) | ES2075900T3 (en) |
| FI (1) | FI104233B (en) |
| NO (1) | NO301577B1 (en) |
| WO (1) | WO1991001143A1 (en) |
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| US6406696B1 (en) | 1989-10-27 | 2002-06-18 | Tolerance Therapeutics, Inc. | Methods of stimulating the immune system with anti-CD3 antibodies |
| WO1991006319A1 (en) * | 1989-10-27 | 1991-05-16 | Arch Development Corporation | Methods and compositions for promoting immunopotentiation |
| GR1000881B (en) * | 1990-08-06 | 1993-03-16 | Praxis Biolog Inc | Stable vaccine compositions containing interleukins |
| CA2098726A1 (en) * | 1990-12-19 | 1992-06-20 | Edward J. Rozhon | Use of il-4 to enhance immune response to immunogens in vaccines |
| HUT67159A (en) * | 1991-05-13 | 1995-02-28 | Univ California | Liposomal polysaccharide vaccines |
| DE4126983A1 (en) * | 1991-08-15 | 1993-02-18 | Boehringer Mannheim Gmbh | METHOD FOR THE PRODUCTION OF HUMAN-PROTEIN-CONTAINING, PRESERVED MEDICAMENTS FOR INFUSION OR INJECTION USE |
| CA2140878C (en) * | 1992-08-10 | 2007-12-04 | Michael J. Lenardo | Interleukin-4 stimulated t lymphocyte cell death |
| ZA936095B (en) * | 1992-08-21 | 1994-03-14 | Univ Melbourne | Cytokine applications. |
| US6491916B1 (en) | 1994-06-01 | 2002-12-10 | Tolerance Therapeutics, Inc. | Methods and materials for modulation of the immunosuppresive activity and toxicity of monoclonal antibodies |
| WO1996007102A1 (en) * | 1994-09-01 | 1996-03-07 | Wisconsin Alumni Research Foundation | Therapeutic remodeling in aids |
| US6509313B1 (en) | 1996-02-28 | 2003-01-21 | Cornell Research Foundation, Inc. | Stimulation of immune response with low doses of cytokines |
| AU724743B2 (en) * | 1996-05-31 | 2000-09-28 | Genetics Institute, Llc | IL-12 as an adjuvant for Bordetella Pertussis vaccines |
| TW586934B (en) * | 1997-05-19 | 2004-05-11 | Sumitomo Pharma | Immunopotentiating composition |
| US6270758B1 (en) | 1998-10-08 | 2001-08-07 | Duke University | Substantially non-toxic biologically active mucosal adjuvants in vertebrate subjects |
| US6027731A (en) * | 1998-11-17 | 2000-02-22 | Wisconsin Alumni Research Foundation | Pertussis toxin induced lymphocytosis |
| IT1302534B1 (en) | 1998-12-21 | 2000-09-05 | Fidia Advanced Biopolymers Srl | INJECTABLE, BIOCOMPATIBLE AND BIODEGRADABLE COMPOSITIONS INCLUDING AT LEAST A DERIVATIVE OF HYALURONIC ACID, CHONDROGENIC CELLS, FOR |
| SK782002A3 (en) * | 1999-07-21 | 2003-08-05 | Lexigen Pharm Corp | FC fusion proteins for enhancing the immunogenicity of protein and peptide antigens |
| AU1813701A (en) | 1999-12-03 | 2001-06-12 | Celltech Chiroscience Limited | Interleukin-1 muteins useful as vaccine adjuvants |
| US8663634B2 (en) | 2005-07-11 | 2014-03-04 | Macrogenics, Inc. | Methods for the treatment of autoimmune disorders using immunosuppressive monoclonal antibodies with reduced toxicity |
| EP1928418B1 (en) | 2005-09-30 | 2011-12-21 | Lipoxen Technologies Limited | Liposomal vaccine compositions comprising a polysaccharide antigen and a protein adjuvant |
| CA2655080A1 (en) | 2006-06-14 | 2007-12-21 | Macrogenics, Inc. | Methods for the treatment of autoimmune disorders using monoclonal antibodies with reduced toxicity |
| US7709010B2 (en) * | 2007-03-09 | 2010-05-04 | Merck Sharp & Dohme Corp. | Papillomavirus vaccine compositions |
| EP2925297B1 (en) * | 2012-11-02 | 2017-06-21 | CyTuVax | Composition comprising cytokine macro-aggregates |
| JOP20190242A1 (en) * | 2018-10-12 | 2020-04-12 | Serum Institute Of India Pvt Ltd | Combination vaccine composition comprising reduced dose inactivated poliovirus and method for preparing the same |
| WO2020102728A1 (en) | 2018-11-16 | 2020-05-22 | Neoimmunetech, Inc. | Method of treating a tumor with a combination of il-7 protein and an immune checkpoint inhibitor |
| EP4090430A1 (en) | 2020-01-13 | 2022-11-23 | Neoimmune Tech, Inc. | Method of treating a tumor with a combination of il-7 protein and a bispecific antibody |
| US20230210952A1 (en) | 2020-02-05 | 2023-07-06 | Washington University | Method of treating a solid tumor with a combination of an il-7 protein and car-bearing immune cells |
| KR20230098201A (en) | 2020-10-26 | 2023-07-03 | 네오이뮨텍, 인코퍼레이티드 | Methods of inducing stem cell mobilization |
| KR20230104176A (en) | 2020-11-02 | 2023-07-07 | 네오이뮨텍, 인코퍼레이티드 | Use of interleukin-7 for treatment of coronavirus |
| KR20230104175A (en) | 2020-11-05 | 2023-07-07 | 네오이뮨텍, 인코퍼레이티드 | Methods of Treating Tumors Using Combinations of IL-7 Protein and Nucleotide Vaccines |
| KR20240130705A (en) | 2021-12-30 | 2024-08-29 | 네오이뮨텍, 인코퍼레이티드 | Method for treating tumors with a combination of IL-7 protein and VEGF antagonist |
| WO2024102722A1 (en) | 2022-11-07 | 2024-05-16 | Neoimmunetech, Inc. | Methods of treating a tumor with an unmethylated mgmt promoter |
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| JP2849632B2 (en) * | 1988-04-08 | 1999-01-20 | 社団法人北里研究所 | Vaccine preparation |
| DE3818054C2 (en) * | 1988-05-27 | 1994-02-17 | Biotest Pharma Gmbh | Use of a combination of an antigen or a vaccine and human interleukin for the treatment of non-responsiveness to immunological defect states |
| JPH0236129A (en) * | 1988-07-22 | 1990-02-06 | Ajinomoto Co Inc | Agent forpromoting effect of vaccine |
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1990
- 1990-07-16 JP JP51060090A patent/JP3485184B2/en not_active Expired - Fee Related
- 1990-07-16 DE DE69018990T patent/DE69018990T2/en not_active Expired - Fee Related
- 1990-07-16 EP EP90911344A patent/EP0482076B1/en not_active Expired - Lifetime
- 1990-07-16 CA CA002063587A patent/CA2063587C/en not_active Expired - Fee Related
- 1990-07-16 ES ES90911344T patent/ES2075900T3/en not_active Expired - Lifetime
- 1990-07-16 AT AT90911344T patent/ATE121629T1/en not_active IP Right Cessation
- 1990-07-16 DK DK90911344.1T patent/DK0482076T3/en active
- 1990-07-16 KR KR1019920700086A patent/KR0177179B1/en not_active IP Right Cessation
- 1990-07-16 AU AU60500/90A patent/AU648509B2/en not_active Ceased
- 1990-07-16 WO PCT/US1990/003982 patent/WO1991001143A1/en active IP Right Grant
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1992
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- 1992-01-13 FI FI920132A patent/FI104233B/en not_active IP Right Cessation
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| AU648509B2 (en) | 1994-04-28 |
| JP2004002463A (en) | 2004-01-08 |
| EP0482076B1 (en) | 1995-04-26 |
| FI104233B1 (en) | 1999-12-15 |
| KR0177179B1 (en) | 1999-03-20 |
| DE69018990T2 (en) | 1995-12-14 |
| CA2063587A1 (en) | 1991-01-15 |
| ATE121629T1 (en) | 1995-05-15 |
| DE69018990D1 (en) | 1995-06-01 |
| WO1991001143A1 (en) | 1991-02-07 |
| FI104233B (en) | 1999-12-15 |
| AU6050090A (en) | 1991-02-22 |
| FI920132A0 (en) | 1992-01-13 |
| NO920161L (en) | 1992-03-05 |
| JPH04506663A (en) | 1992-11-19 |
| NO920161D0 (en) | 1992-01-13 |
| KR920703103A (en) | 1992-12-17 |
| CA2063587C (en) | 2005-08-30 |
| NO301577B1 (en) | 1997-11-17 |
| JP3485184B2 (en) | 2004-01-13 |
| ES2075900T3 (en) | 1995-10-16 |
| DK0482076T3 (en) | 1995-07-17 |
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